The Influence of Endogenous Hormones on Culture Capability of Different Explants in Rice

The endogenous hormones (EHs) content of different explants (anther, young panicle, young embryo and mature embryo) and calli with different culture capability were analyzed by means of high performance liquid chromatography (HPLC). The results showed that the contents and ratio of endogenous hormones were one of the key factors affecting callus induction frequencies (CIF) and green plantlet differentiation frequencies (GPDF). The influence of endogenous hormones of different explants on CIF represented as: Zoatin ribosile (ZR) showed negative effect, indole-3-acetic acid (IAA) did positive effect, and gibberellic acid (GA) did negative effect except for mature embryos. The influence of endogenous hormones on green plantlet differentiation frequency (GPDF) showed: IAA and GA were negative effect; abscisic acid (ABA) and zeatin+ zeatin riboside (Z+ZR) were positive effect. The mixture ratio of endogenous hormones played a role on CIF and GPDF. IAA/Z+ZR had a positive effect on CIF, and there was a notable positive correlation between Z+ZR/ IAA and GPDF, so was between ABA/IAA and GPDF.

Tissue Culture Responses from Different Explants of Rice

Different culture explants, including anther, young panicle, young embryo, and mature embryo, from 19 rice varieties were used for callus induction and green plantlet differentiation. The culture efficiency differed significantly among the four types of explants, and varied from genotype to genotype. Callus induction frequency presented significantly positive correlation each between anther and young panicle, anther and mature embryo, and young panicle and young embryo. Green plantlet differentiation showed no relationship between different types of explants. In addition, no relationship was found between callus induction frequency and green plantlet differentiation frequency.

Study on Initial Explants Induction of Litsea cubeba

[ Objectlve] The aim of this study was to explore an explants induction method for L. cubeba. [ Method ] Different types of L. cubeba stem segments were collected at different time as explants materials for induction and culture. [ Result ] The results showed that sterilization with 0. 1% HgC12 for 10 min achieved better effects, with an explants contamination rate of dO. 0% and a mortality rate of 15.0% ; stem segments of the 2na -5th buds below the terminal bud were better explants materials for initial induction; the best sampling time was January, with an induction rate as high as 81.1%, while the contamination rate was only 11.7% ; modified MS ＋ 1.0 mg/L of 6-BA ＋ 0.2 mg/L of NAA was the optimal medium for explants induction of L. cubeba. [ Conclusion] Semi-lignified stem segments of L cubeba collected in January, sterilized with 0.1% HgCI2 for 10 min and cultured in modified MS ＋ 1.0 mg/L of 6-BA ＋ 0.2 mg/L of NAA a- chieved better induction effects on L. cubeba seedlings.

Influences of Genotype and Explant on Callus Induction and Shoot Regeneration in Sunflower （Helianthus annuus L.）

In vitro regeneration was performed with the aim of developing efficient callus and shoot regeneration from different explants of sunflower （Helianthus annuus L.）. The 6 genotypes （RHAI0, RHA 14, RHA15, PR6404, N Record 109/Sanay 3-5, N Record 109/Sera） were used as plant materials. The roots, hypocotyls and cotyledons were excised from 4 day-old seedlings and cultured on embryo induction medium （EIM） supplemented with Benzylaminopurine （BAP）, Naphthaleneacetic acid （NAA） and Gibberellic acid （GA3）. The experiments were kept in 18/6 hour light/dark photoperiod at 26± 2 ℃for four weeks. The rates of callus and root organ formation on explants were 67%-100% and 7%-31% respectively, depending on the genotype. Root explants produced statistically high callus formation （2.6 score） compared to cotyledon （2.0 score） and hypocotyl explants （2.5 score） at all sunflower genotypes used. The highest shoot regeneration was obtained from RHAI 5 （7%） while PR6404 （100%） produced the highest callus formation.

Tissue Culture Nursery Technology of Curcuma alismatifolia‘Kimono Rose’

[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we identified the most optimal explants and optimized the culture conditions for cluster buds induction and proliferation.This was achieved by incorporating MS medium with varying concentrations of 6-BA and NAA,thereby establishing a foundation for the large-scale production of C.alismatifolia tissue culture seedlings.[Results]The optimal explant for C.alismatifolia was identified as a lateral bud.The most effective cluster buds induction medium was determined to be MS+6-BA 5 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.The optimal cluster buds proliferation medium was found to be MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.[Conclusions]The findings of this study can provide a foundation for the enhancement of the industrialized breeding system of tissue culture propagation of C.alismatifolia.

Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L.(Scrophulariaceae)-an ethnomedicinal herb

Objective:To develop an improved protocol for micropropagation of ethnomedicinally important Scoparia dulcis(S.dulcis) L.Methods:Explants were inoculated on MS basal medium supplemented with kinelin and 6-benzylaminopurine for shoot bud induction.To enhance the shoot induction,various auxins like 3-indoleacetic acid or 3-indolebutyric acid or a-naphthylacetic acid were tested along with 2.32 M KI and 4.44 μ M BAP.The regenerated shoots were rooted in half strength MS medium supplemented with various concentrations of IAA,IBA or NAA.After roots were developed,the plantlets were transplanted to pots filled with vermiculate and sand and kept in growth chamber with 70%-80%humidity under 16 h photoperiod.After acclimatization,the plantlets were transferred to the garden and survival percentage was calculated.Data were statistically analyzed and means were compared using Duncan's multiple range test(P<0.05).Results:An in vitro method was developed to induce high frequency shoots regeneration from stem,mature leaf and young leaf explants of S.dulcis.Shoot induction on young leaf explants was most successful in MS medium supplemented with combination of two cytokinins(2.32 μ M KI and 4.44 μ M BAP) 2.85 μ M IAA,10%CM and 1 483.79 μM adenine sulfate.A single young leaf explant was capable of producing 59 shoots after 13 days of culture. Flower was induced in medium supplemented with combination of KI and BAP.Conclusions: Cytokinins are the key factor to induce the direct shoot regeneration and flowering of S.dulcis.

Tissue Culture of Red Sandalwood (Pterocarpus santalinus)

[Objectives] This study was conducted to develop a rapid propagation method for red sandalwood ( Pterocarpus santalinus ) by tissue culture.[Methods] The well-grown red sandalwood seed embryos were inoculated into three kinds of culture media after aseptic treatment,and the aseptic explants were obtained and inoculated into six kinds of media for light culture.[Results] In the best disinfection schemes of red sandalwood,disinfecting with HgCl 2 for 8 min achieved the highest germination and survival rates;when the medium for inducing red sandalwood explants was MS+0.2 mg/L IBA,the induction rate reached a maximum value;and when the culture medium for inducing stem segments of aseptic red sandalwood plantlets was MS+3.0 mg/L 6-BA+0.3 mg/L IBA,the growth of the stem segments achieved the best effect.The optimal medium for inducing red sandalwood explants was MS+IBA 0.2 mg/L,and the optimal medium for inducing stem segments of red sandalwood was MS+3.0 mg/L 6-BA+0.3 mg/L IBA.[Conclusions] The results of this study have a large reproductive coefficient,simple process and low cost,which have outstanding value for promoting the breeding and promotion of red sandalwood seedlings.

Study on High-frequency Callus Induction From Aseptic Plantlets of Anthuium andraeanum

[Objectives]This study was conducted to improve the efficiency of callus induction and redifferentiation,and construct high-frequency plant regeneration techniques of tissue culture in Anthuium andraeanum.[Methods]The effects of different genotypes,explant types and hormonal conditions on callus induction and re-differentiation of A.andraeanum were studied by using the aseptic A.andraeanum test-tube plantlets as test materials.[Results]Among the four kinds of aseptic A.andraeanum plantlets,the callus induction using stem segments with leaves was the best,followed by stem segments and leaves,and the petioles were the worst;among the six A.andraeanum varieties tested,the callus production rates of four varieties reached 100%;and the callus differentiation rate reached 93.3%-100%through the organogenesis pathway,and the suitable differentiation medium was 1/2MS+ZT 0.5 mg/L+2,4-D 0.1 mg/L.[Conclusions]The research results provide a new experimental basis for optimizing the technical system of A.andraeanum rapid propagation.

硅藻抑制剂二氧化锗在海带体细胞诱导体系中的影响

为探究硅藻抑制剂二氧化锗在诱导体系中对硅藻的抑制作用及对诱导体系的影响,以海带幼孢子体为外植体,在2%PESI培养基中分别加入质量浓度为0、1.00、2.00、2.50、3.00、3.50 mg/L和4.00 mg/L的二氧化锗,探讨不同质量浓度二氧化锗对诱导体系中海带透明细丝发生率及愈伤组织诱导率的影响。试验结果显示:不同质量浓度二氧化锗对海带体细胞诱导体系的影响极显著(P<0.01);体系诱导至第60天时,随二氧化锗质量浓度的提高,透明细丝的细胞组成数及分支逐渐减少,3.50 mg/L及以上组几乎不出现透明细丝;透明细丝发生率在低质量浓度组(0~2.50 mg/L)差异不显著,均超过80%,显著高于高质量浓度组(P<0.05);第80天时,愈伤组织诱导率在1.00、2.00 mg/L低质量浓度组差异不显著,分别为83.33%、73.61%,显著高于高质量浓度组(P<0.05);应用1.00 mg/L的二氧化锗对已污染硅藻的诱导体系进行初步治疗,抑制硅藻污染效果显著。综上所述,在海带体细胞诱导体系中加入二氧化锗的最佳质量浓度为1.00 mg/L,在不影响海带体细胞诱导体系的前提下,可有效抑制硅藻污染。

苹果砧木新品种鲁砧1号离体叶片高效不定植株再生

【目的】建立苹果半矮化砧木新品种鲁砧1号离体叶片高效不定梢再生技术体系,为该砧木快繁和遗传改良奠定技术基础。【方法】以鲁砧1号无菌苗离体叶片为外植体,研究碳源、细胞分裂素种类和质量浓度对叶片不定梢再生的影响;以不定梢为试材,研究基本培养基、蔗糖质量浓度对不定梢生根的影响。【结果】MS添加较低质量浓度(0.6 mg·L^(-1))细胞分裂素(TDZ)时,碳源物质为D-山梨醇的叶片不定梢再生率显著高于蔗糖;其他激素处理下,D-山梨醇和蔗糖之间叶片不定梢再生率无显著差异。两种碳源上的平均每叶不定芽(梢)数,都表现较高细胞分裂素质量浓度处理高于或显著高于较低细胞分裂素质量浓度处理,以TDZ质量浓度为1 mg·L^(-1)时产生的不定芽数最多,平均每叶不定芽数为3.8~4个。在不定芽诱导培养基上,6-苄基氨基嘌呤(BA)比TDZ更容易诱导产生直接伸长生长的不定梢。不定梢生根率和平均每株生根条数,两种基本培养基及两个蔗糖质量浓度之间都表现差异不显著,但以1/4MS基本培养基和20 g·L^(-1)蔗糖组合的生根培养基上获得的生根率和单株生根条数最高,分别超过93%和5.8。【结论】鲁砧1号离体叶片容易诱导再生不定芽和不定梢生根,诱导叶片不定芽再生最佳培养基为MS添加1 mg·L^(-1)TDZ、0.3 mg·L^(-1)IBA和30 g·L^(-1)蔗糖,最佳生根培养基为1/4MS添加0.3~0.5 mg·L^(-1)IBA和20 g·L^(-1)蔗糖。

‘无核白’葡萄多倍体的化学诱导及鉴定

以‘无核白’葡萄分生愈伤组织和胚性愈伤组织为外植体材料,分别通过秋水仙素与安磺灵浸渍法进行诱导,比较不同外植体材料成苗后的存活率和诱变率差异,利用流式细胞仪鉴定成苗倍性。结果显示,秋水仙素浸渍法诱导‘无核白’分生愈伤组织,分别在浓度为100 mg·L^(-1)处理48 h和250 mg·L^(-1)处理24 h获得嵌合体苗,前者诱变率为2%,后者诱变率为4%,且经组培方式纯化获得纯合四倍体;利用安磺灵浸渍法诱导‘无核白’分生愈伤组织获得了7株嵌合体苗,1.73 mg·L^(-1)处理48 h的试验组获得最高存活率为80%,1.73 mg·L^(-1)处理72 h的试验组获得最高诱变率为8.0%;采用秋水仙素浸渍法对‘无核白’胚性愈伤组织进行加倍处理,未获得成苗;采用安磺灵浸渍法对‘无核白’胚性愈伤组织进行加倍处理,当处理浓度及时间为5.19 mg·L^(-1)、24 h时,获得成苗4株,其中2株为嵌合体。

不同激素配比对矩镰荚苜蓿4种外植体愈伤组织诱导的影响

【目的】筛选出矩镰荚苜蓿(Medicago archiducis-nicolai)最佳外植体及最佳再生条件。【方法】以矩镰荚苜蓿种子为材料,选择MS(含0.8%琼脂+3%蔗糖)为基本培养基,添加不同种类及浓度配比生长调节剂对矩镰荚苜蓿子叶、叶柄、茎、叶片进行诱导愈伤和愈伤再分化。【结果】外植体诱导率由高到低依次为:茎>叶柄>真叶>子叶,外植体褐化率由高到低依次为:茎<真叶<叶柄<子叶;在1.5 mg/L 2,4-D+0.1 mg/L KT+MS+0.8%琼脂+3%蔗糖条件下最适合真叶诱导愈伤,诱导率为78%,褐化率为8%;1.5 mg/L 2,4-D+1.0 mg/L KT+MS+0.8%琼脂+3%蔗糖最适合子叶、叶柄、茎诱导愈伤,在此培养基中,茎的出愈率为96%,褐化率为0,叶柄的出愈率最高,为92%,褐化率为2%,子叶的出愈率为60%,褐化率为4%;在0.3 mg/L NAA+1.0 mg/L KT+MS+0.8%琼脂+3%蔗糖条件下最适合子叶、叶柄诱导胚性愈伤;在0.3 mg/L NAA+1.0 mg/L 6-BA+MS+0.8%琼脂+3%蔗糖条件下最适合真叶诱导胚性愈伤;在0.3 mg/L NAA+0.5 mg/L KT+MS+0.8%琼脂+3%蔗糖条件下最适合茎诱导胚性愈伤。【结论】4种外植体均产生愈伤组织,且进一步诱导愈伤组织可分化形成体细胞胚。4种外植体中,茎的愈伤诱导率最高,褐化率最低,为最佳外植体。

马蹄金愈伤组织诱导及植株再生研究

以马蹄金无菌苗的下胚轴、子叶和叶片作为外植体 ,结合不同植物生长调节剂组合和不同基本培养基 ,探讨了马蹄金离体培养的方法和影响因素。结果表明 ,不同植物生长调节剂组合的培养基对愈伤组织诱导频率有显著影响 ,以MS培养基作愈伤组织诱导的基本培养基效果较好 ,使用MS +1.0mg/L 2 ,4 -D +0 .2mg/L6 -BA或MS +1.0mg/L 2 ,4 -D +0 .2mg/LKT作为愈伤组织诱导培养基 ,对 3种外植体愈伤组织诱导的效果均较好 ,出愈率均为 10 0 % ,而且愈伤组织质地好、生长旺盛 ,再用MS +0 .1mg/L 2 ,4 -D +2 .0mg/L 6-BA +3.0mg/LAgNO3 作愈伤组织分化培养基出苗效果最佳 ,愈伤组织分化频率达 2 0 .7%。

玉米愈伤组织再生体系的研究

用玉米胚性愈伤组织作为转化受体时,不同基因型在相同2,4-D浓度的培养基上诱导,其诱导率有较大差别,齐319最高;胚龄对诱导率的影响也较大,10d左右最易诱导,过大或过小均不易诱导胚性愈伤;培养基中2,4-D的浓度对玉米愈伤组织的诱导率有极大影响,不同基因型的最大胚性愈伤组织诱导率所需的2,4-D浓度也不相同,一般为0.5～1.0mg/L。愈伤组织分化时不同基因型所需最适6-BA浓度不同,齐319、515的愈伤组织最适合分化成苗的6-BA浓度为1mg/L,鲁原341愈伤组织最适6-BA浓度为0.5mg/L。

不同外植体和激素对银杏愈伤组织诱导和生长的影响

以银杏胚的不同部位子叶、胚芽、胚轴、胚根为外植体,比较不同外源激素和培养条件对银杏愈伤组织诱导和继代培养的影响,结果表明子叶最易诱导愈伤组织,且在暗条件下诱导效果最好,2,4-D和BA影响子叶的愈伤诱导能力;以茎段为外植体,发现生长素为茎段愈伤组织诱导所必需,且NAA诱导效果好于2,4-D;以叶片为外植体,发现生长素中的NAA诱导愈伤效果最好,雌雄叶的诱导能力相同.在愈伤组织生长过程中,NAA是子叶和茎段愈伤组织继代培养必需的,BA对茎段愈伤组织生长速度无显著影响,但可使愈伤组织致密,并有利于分化.MS+1mg/LNAA+0.005mg/LKT(或BA)的培养基对叶片愈伤组织生长最理想,雄叶愈伤生长较快,光照培养促进愈伤组织生长.

高粱不同外植体离体培养

不同材料愈伤组织诱导培养结果表明，幼穗、茎尖和成熟胚的最高出愈率达１００％，子粒达９３％，幼胚仅为７４％。幼穗不但出愈率高，且愈伤组织胚性状况好，分化出苗率高。幼胚出愈率虽低，但其分化率明显高于茎尖、成熟胚和子粒的分化率。

无籽西瓜种苗愈伤组织诱导研究

用不同基因型无籽西瓜进行了愈伤组织诱导研究.结果表明:无籽西瓜种苗的最佳愈伤组织诱导培养基组合是MS+NAA1 0mg·L-1+6 BA0 5mg·L-1(C5培养基);种苗下胚轴和子叶是适宜外植体;外植体愈伤组织的产生首先需要黑暗条件的启动,其扩增与生长分化则须在光照条件下进行;不同基因型外植体在C5培养基上的愈伤组织诱导率和诱导指数存在显著差异(P=0 05水平).

不同蓝莓基因型的组织培养技术研究

以南高丛蓝莓和免眼蓝莓当年生半木质化枝条做外植体，通过对不同天气的外植体采集、外植体单株芽数的对比、灭菌时阊的不同处理、不同激素培养基培养的对比试验，发现其中外植体采集以晴天，叶片无水珠采集为宜，以规模化生产为目的的外植体以单株2个芽为宜，外植体最佳灭菌方法o．1％升汞，嫩部处理6min，木质化程度相对较高部分处理10min，污染率在27％以下。芽诱导培养基配方以MWPM＋ZT（2．0—3．0mg／L）＋NAA0．1mg,／L，琼脂6．0g／L，蔗糖25g／L，pH5．0为宜，平均萌芽时间为8d，平均萌芽率在80％以上，且芽质量较好。

Cu^(2+)对小麦幼胚脱分化和高频再生特性的影响

以小麦品种(系)千斤早和9848的幼胚组织为材料,研究了Cu2+对小麦幼胚组织脱分化和再分化特性的影响。结果表明,诱导培养基中的Cu2+对小麦幼胚愈伤组织的形成无显著影响,但可显著提高被诱导愈伤组织的再分化能力。在供试Cu2+浓度范围内,千斤早的愈伤组织再分化率比对照提高了37.9%～72.4%,9848比对照提高了8.2%～21.2%。分化培养基中的Cu2+对小麦幼胚愈伤组织再分化的影响表现出低浓度促进、高浓度抑制的效应,对愈伤组织再分化的最佳正向效应浓度千斤早为1.0μmol/L,比对照提高101.4%;9848为1.0～5.0μmol/L,分别比对照提高102.5%～116.6%;Cu2+对小麦幼胚愈伤组织再分化的影响远高于对幼胚组织脱分化的影响。

无患子愈伤组织诱导的多因子正交试验研究

目的：探寻无患子愈伤组织诱导的最佳外植体、培养基和培养条件.方法：采用5因素混合水平正交设计,用DPS2000统计软件对其结果进行方差分析和多重比较（SSR）.结果：诱导愈伤组织的最佳外植体为当年生枝条水培芽的嫩茎段,用0.2%HgCl2消毒2min,其无菌存活率为98.33%,培养条件为光照或黑暗,最佳培养基配方为：MS（B5）＋6-BA 1.5mg·l^-1＋NAA0.2mg·l^-1,其愈伤组织的诱导率为97.78%.结论：激素配比是影响愈伤组织诱导的主要因素.较高的6-BA浓度（1.5～2.5 mg·l^-1）与较低的NAA浓度（0.2mg·l^-1）配比,有利于无患子愈伤组织的诱导.